Effects of cyclosporin A on the course of murine alveolar echinococcosis and on specific cellular and humoral immune responses against Echinococcus multilocularis.

The effects of cyclosporin A (CsA) on Echinococcus multilocularis (E. multilocularis) metacestode growth, and on the specific immune responses of the hosts, were examined in AKR mice. Mice were intra-peritoneally infected with a metacestode homogenate. CsA (40 mg kg(-1) day(-1)) was injected subcutaneously from the 45th day after infection (Group 1), and from the day before infection (Group 2) until the day of autopsy (days 125 and 80, respectively). Results showed that unlike ths situation with some other helminthiases, CsA had no antiparasitic effect, although it lengthened the maturation time of protoscoleces in Group 1. The parasitic burden, unmodified in Group 1, was significantly enhanced in Group 2. This enhancement was associated with a decrease in antibody levels, whereas the delayed-type hypersensitivity was decreased in the two groups. These results confirm the role of cellular immunity in controlling the first stages of the larval development of E. multilocularis and indicate the necessity for a careful follow-up of any recurrence of alveolar echinococcosis in patients treated with CsA after liver transplantation.

Seroepidemiologic screening of Echinococcus multilocularis infection in a European area endemic for alveolar echinococcosis.

In a serologic survey for Echinococcus multilocularis infection, we screened sera from 7,884 subjects from the Doubs Departement in France, an area endemic for alveolar echinococcosis (AE) of the liver. An enzyme-linked immunosorbent assay (ELISA) with a highly species-specific antigen (Em2) and an E. multilocularis crude antigen (Emc) was used for screening. An evaluation of the cost/benefit relationship of this screening, followed by therapeutic management of patients, was made and compared with the actual cost of the follow-up and treatment of the disease in symptomatic cases in this endemic area. Antibody reactions to Em2 and/or Emc made possible the detection of eight asymptomatic clinical cases (seroprevalence averaging 1/1,000), with typical lesions of active AE revealed by abdominal ultrasonography and computed tomography. All were seropositive using the Emc ELISA but two were seronegative using the Em2 ELISA. In five additional seropositive cases, the radiologic investigations revealed small calcified lesions similar to the lesions of abortive AE previously found in Alaska. The cost of this serologic screening program per screened subject and per diagnosed case averaged 50.00 French Francs (FF) (U.S. $8.60) and 60,000.00 FF (U.S. $10,909.00), respectively. The cost of diagnosis, follow-up and treatment of the patients was 5,086.00 FF (U.S. $929.00) per patient per month in the case of diseases diagnosed by the screening program and 7,086.00 FF (U.S. $1,288.00) per patient per month for patients with symptomatic AE. This survey indicates a high prevalence of AE in the target area; it confirms the long latency period of the larval growth in human AE and shows that abortive AE is present in Europe. The use of both the Emc and Em2 ELISAs seems to be better than using the Em2 ELISA alone. The cost of the hospitalization and treatment of the eight screened patients would appear to be relatively high. Even though two of them were asymptomatic, they had very severe forms of the disease. In fact, the total cost was much lower than the actual cost of the disease when diagnosed from clinical symptoms.

Immunocytochemical evidence that the same neurons in the human infundibular nucleus are stained with anti-endorphins and antisera of other related peptides.

Neurons immunoreactive with anti-beta-endorphin can be detected in the infundibular region of human fetal hypothalami from the 11th week of development. In the human adult, neurons stained with anti-beta-endorphin are located in the infundibular nucleus and are also stained with anti(17-39) ACTH. Their fibers terminate in contact with non/immunoreactive perikarya or close to vessels of the pituitary stalk. The neurons most reactive with anti(17-39)ACTH or anti-beta-endorphin are also immunoreactive with anti-alpha-MSH, anti-beta-MSH, anti-alpha-endorphin, anti(1-24)ACTH or anti-beta-LPH. These results suggest that a molecule similar to pro-opiocortin of fragment(s) of it is secreted by these neurons.

Coexistence of acetylcholinesterase-, human growth hormone-releasing factor(1-37)-, alpha-melanotropin- and melanin-concentrating hormone-like immunoreactivities in neurons of the rat hypothalamus: a light and electron microscope study.

Using an antiserum (AS) raised against rat cerebral acetylcholinesterase (AChE), we revealed a neuron population in lateral and dorsal areas of the posterior rat hypothalamus. These neurons were previously described using antibodies to human growth hormone-releasing factor(1-37) (GRF-37), alpha-melanotropin (alpha-MSH) and melanin-concentrating hormone (MCH). Different intracytoplasmic distributions of the immunodeposits were observed depending on the used serum. Ultrastructural investigations demonstrated that AChE-AS labeled rough endoplasmic reticulum and nuclear envelope in control rats. MCH-AS stained Golgi apparatus in control animals and secretory granules in colchicine-injected rats. GRF-37-AS always revealed secretory granules, and alpha-MSH-AS gave the same staining only after colchicine injection.

Comparative study of the neuronal populations containing beta-endorphin, corticotropin and dopamine in the arcuate nucleus of the rat hypothalamus.

Neurons simultaneously immunoreactive with anti-beta-endorphin and anti-(17-39) corticotropin (anti(17-39) ACTH) have been detected in the arcuate and ventromedial nuclei of the hypothalamus of colchicine-treated rats. These neurons are different from those fluorescent with the Falck and Hillarp technique. These results show that in the arcuate nucleus, immunoreactive ACTH and beta-endorphine are stored in the same neurons which are different from dopamine-containing neurons.

Analysis of immunoglobulin G subclass in the serum antibody responses of alveolar echinococcosis patients after surgical treatment and chemotherapy as an aid to assessing the outcome.

Immunoglobulin G (IgG) subclass-specific antibody responses were evaluated for the follow-up of alveolar echinococcosis (AE) patients. Seventy-four sequentially collected sera from 25 Chinese and French AE cases who underwent surgery including hepatectomy, liver transplant and/or chemotherapy were analysed quantitatively and qualitatively during the clinical follow-up period. These AE patients were classified in 4 groups--cured, improved, stabilized, or aggravated. Serum antibody levels of the subclasses IgG1 and IgG4 were significantly higher in the AE patients than in healthy controls. IgG1 and IgG4 isotypes in AE patients were the most sensitive IgG antibody response in an enzyme-linked immunosorbent assay (ELISA) and in binding to antigens of 44kDa, 35kDa, 21kDa and 17.5kDa in an Echinococcus multilocularis protoscolex extract after Western blotting. In AE cases classed as cured or improved, IgG subclass antibody levels tended to decrease earlier than total IgG levels, especially IgG4 antibody levels which became negative within one year after successful treatment. IgG4 antibody levels also decreased in most of the improved cases. Increasing or unchanged levels of IgG4 and IgG1 antibodies were demonstrated in both stabilized and aggravated AE cases using both ELISA and immunoblot assays. Reappearance of specific IgG4 antibodies was a strong indication of recurrence, especially in liver transplant patients. Combined quantitative and qualitative assessment of IgG1 and IgG4 antibodies may be potentially useful for the serological follow-up of human AE.

Soluble IL-2-receptor and CD8 in the serum and the periparasitic granuloma of patients with alveolar echinococcosis.

Cellular immunity plays a key role in the defence against the larva of the cestode Echinococcus multilocularis. This larva is responsible for alveolar echinococcosis (AE) of the liver, a rare parasitic disease which occurs in endemic areas including European alpine countries, Alaska, the USSR, Western China and Northern Japan. We have shown a marked decrease of the CD8+ T-cell population in the blood and we have described an infiltrate composed mainly of activated CD8+ T-cells in the liver lesions of most patients with AE. In this study, we assessed the serum level of soluble IL-2-receptor (sIL-2R) and CD8 (sCD8) in 37 patients (23 men, 14 women, mean age 59.5 yrs) with a histologically proven AE. The results, obtained using sandwich ELISA, were compared to those of healthy controls and correlated to parameters evaluating the severity of the disease. The mean serum levels of sIL-2R were significantly higher in AE patients than in controls. There was a significant correlation between sIL-2R levels and both the volume of parasitic lesions and a calculated index of severity of the disease. The mean serum levels of sCD8 did not differ significantly from the values obtained in controls. These results indicate that the infiltration of the liver by CD8+ T-lymphocytes is not associated with an increased release of sCD8 into the serum. The circulating levels of sIL-2R appear to reflect the extent as well as the severity of the disease. Immunostaining of the cells of the periparasitic granuloma suggests that the cell origin of the sIL-2R could be macrophages rather than T-lymphocytes.

A twenty-year history of alveolar echinococcosis: analysis of a series of 117 patients from eastern France.

OBJECTIVES: Alveolar echinococcosis of the liver is a very rare and severe parasitic disease due to the growth of the larva of Echinococcus multilocularis. The aim of this paper was to describe a 20-year study of the epidemiological, clinical and therapeutic aspects of alveolar echinococcosis in eastern France. DESIGN: One hundred and seventeen consecutive cases, diagnosed and followed in our liver unit, were studied from 1972 to 1993. METHODS: Data from 85 patients followed since 1983 (period B) were compared to data from a first series of 32 patients (period A) collected from 1972 to 1982; 1983 was chosen as the cut-off year because of the numerous changes that occurred in the diagnosis, follow-up and treatment of the disease at this time, in particular the introduction of parasitostatic benzimidazoles. RESULTS: The results of patient follow-up were evaluated in December 1997. The cumulative prevalence was 2.5 per 100,000 persons in period A whereas it reached 6.6 per 100,000 in period B. The annual incidence in period B was 7.3 on average, compared with 2.7 in period A. Twenty-nine per cent of patients from period B were asymptomatic at the time of diagnosis compared with 10% in period A. This change was correlated with less advanced liver lesions, and was related to the extensive use of abdominal ultrasound, and from 1987, serological screening. Curative resections were performed in 24% of the cases in period B versus only 3% in period A. From 1986, liver transplantations were performed in eight patients from period A and 13 patients from period B. In period B, palliative surgery was frequently replaced by radiological non-operative procedures to treat abscesses and jaundice. From 1982, 73 patients received benzimidazoles for a period of time ranging from 4 to 138 months. Stabilization of the lesions was observed in two-thirds of the patients. Episodes of jaundice or digestive haemorrhage due to portal hypertension were 31.5 and 11 times less frequent respectively in patients from period B compared with period A. Actuarial survival at 5 years improved from 67% in period A to 88% in period B in patients of similar age. CONCLUSIONS: Radical changes in the diagnosis and the management of alveolar echinococcosis have occurred during the last decade. Together they have contributed to an improvement in the status of the patients affected by this very severe parasitic disease.

[Acute allergic colitis caused by ACTH]. / Colite allergique aiguë à l'ACTH.

The case of a woman suffering from urticaria of the colon is reported. The original endoscopic images were described and its allergic (corticotrophin) origin was documented through clinical arguments and in vivo and in vitro tests. The term acute allergic colitis seems to be more suitable taking into account the distribution, the cause and the development of this disease.

Specific cellular and humoral immune responses induced by different antigen preparations of Echinococcus multilocularis metacestodes in patients with alveolar echinococcosis.

A specific proliferation of the peripheral blood mononuclear cells (PBMC) stimulated by antigens of Echinococcus sp. has been shown in patients with cystic as well as alveolar echinococcosis. However, the development of a major granulomatous reaction around the parasitic larvae is a characteristic feature of the local immune response to E. multilocularis while humoral immune responses seem to predominate in E. granulosus infection. The aim of this study was to analyse the specific proliferation of the PBMC from 36 patients with alveolar echinococcosis, and 23 controls, induced by a crude preparation of E. multilocularis (EmcAg) as well as by two E. multilocularis specific antigen preparations, the Em2 antigen and the protoscolex (ESAg) antigen. The significant correlation observed between the proliferation index either by Em2 and ES or by Emc suggests that both antigens account for an important part of the lymphocyte proliferative response. The strong effect of these species specific antigens on lymphocyte proliferation is confirmed by the comparison of the results obtained in this study to those obtained in a previous study of specific cellular immunity to E. granulosus antigens in patients with cystic echinococcosis. The proliferation indices were significantly elevated in all those 7 patients with a proven AE who were sero-negative using the Emc ELISA as well as in the 12 patients also seronegative, but using the Em2 ELISA. In 5 seronegative patients who had had a complete resection of the parasitic lesions two years before the lymphocyte proliferation evaluation, the PI was above the threshold value for two dilutions of EmcAg. PI under the threshold values were obtained only in patients with residual lesions. These results suggest that E. multilocularis specific antigens promote the proliferation of lymphocytes which could be TH1 cells, responsible for the intense periparasitic granulomatous reaction characteristic of alveolar echinococcosis.

[Contribution of immunocytochemistry to the study of the development of neuroglandular peptidergic systems in the human fetal hypothalamus]. / Apport de l'immunocytochimie à l'étude du développement de systèmes neuroglandulaires peptidergiques dans l'hypothalamus foetal humain.

Immunohistochemistry makes possible the in situ detection of neuropeptides in the cell bodies were they are synthesized, in the fibers that carry them, and in endings. Immunohistochemistry appears necessary to identify and map peptidergic neurons and to study their ontogeny. From 1975, we have carried the immunohistochemical study of several hypothalamic neuronal populations in the human fetus: LH-RH (1976), somatostatin (1977), pro-opiocortin (1978), vasopressin and oxytocin (1979), corticoliberin (1982), somatocrinin (1983), and hypothalamic neurons containing an unidentified peptide (1984). Comparative ontogenetical studies have also been performed in rats.

Infundibular neurons of the human hypothalamus simultaneously reactive with antisera against endorphins, ACTH, MSH and beta-LPH.

In man, discrete neurons of the infundibular (arcuate) nucleus contain compounds that can be stained with anti-endorphin (alpha and beta), anti-ACTH, anti-MSH (alpha and beta) and anti-beta-LPH immune sera (I.S.). In the fetus, certain neurons stain with anti-beta-endorphin or anti((17--39)ACTH starting from the 11th week of fetal life. At the ultrastructural level, these neurons contain elementary granules that are immunoreactive with anti-beta-endorphin. In the adult, neurons immunoreactive with anti-beta-endorphin are found in the infundibular nucleus. Their axonal fibers terminate around blood vessels in the neurovascular zone and in the pituitary stalk, or establish contacts with non-immunoreactive perikarya of the infundibular nucleus. These neurons can be stained with anti(17--39)ACTH and anti-beta-endorphin I.S. The most reactive are also stained moderately with anti-alpha-MSH, anti-beta-MSH, anti-beta-LPH, anti-alpha-endorphin, or anti(1--24)ACTH I.S. These results indicate that, in man, compound(s) identical with or immunologically related to endorphins, beta-LPH, ACTH and MSH are secreted by certain hypothalamic neurons. These agents probably originate from a common precursor molecula similar to the so-called pro-opiocortin.

[Immunocytochemical study of the maturation of the hypothalamo-neuro-hypophysial axes in the human fetus (author's transl)]. / Etude immunocytologique de la maturation des axes neuroglandulaires hypothalamo-neurohypophysaires chez le foetus humain.

1. The development of the hypothalamo-neurohypophysial system was studied using immunocytochemistry with various antisera : anti-neurophysin, anti-oxytocin, anti-vasopressin, anti-vasotocin and anti-somatostatin. 2. Immunocytochemical investigation shows that anti-vasopressin and anti-vasotocin sera react with both vasopressin and vasotocin, whereas the anti-oxytocin serum specifically reveals the oxytocin-containing structures (Fig. 1g, h, i). 3. Perikarya stained with anti-neurophysin, anti-vasopressin and anti-vasotocin sera can be seen from the 11th week of fetal life (Fig. 1a) first in the supra-optic nucleus (SON), then in the para-ventricular nucleus (PVN). Their axons reach the neural lobe as early as the 11th week (Fig. 1b, c). 4. Oxytocin-containing perikarya appear simultaneously in the PVN (Fig. 1e) and SON during the 13th week. 5. From the 16th week, neurons stained with the anti-somatostatin serum can be seen among the neurophysin-positive cell-bodies of the SON and PVN. 6. The various populations of magnocellular neurons show a significant increase in number, especially up to the 19th week, and an increase in their hormonal content up to birth.

[Antigenic determinants of beta-LPH, beta-MSH, alpha-endorphin, ACTH and alpha-MSH revealed by anti-beta-endorphin in neurons of the human infundibular nucleus]. / Présence de déterminants antigéniques de la beta-LPH, de la beta-MSH, de l'alpha-endorphine, de l'ACTH et de l'alpha-MSH dans les neurones révélés par l'anti-beta-endorphine au niveau du noyau infundibulaire de l'Homme.

Comparison of adjacent serial sections of the tubero-infundibular region of Human adult hypothalamus demonstrates that the same perikarya, axons and terminals are stained both with anti-beta-endorphin and anti 17-39 ACTH antisera. The most immunoreactive of these neurons are also revealed with anti alpha-endorphin, anti alpha and beta-MSH, anti-1-24 ACTH and anti beta-LPH. These results suggest that neurons of the infundibular nucleus can store and probably secrete peptide similar to propiocortin or fragment(s) of this molecule.

[Description of neurons immunoreactive to anti-beta-endorphin immune serum present in the infundibular nucleus in man]. / Description des neurones immunoréactifs à un immunsérum anti beta-endorphine présents dans le noyau infundibulaire chez l'homme.

Neurons containing peptide (s) immunologically related to beta-endorphin have been detected by immunochemistry in human adult and fetal hypothalami. Their perikarya are located in the infundibular nucleus. Some fibres terminate close to vessels in the median eminence, others form pericellular baskets around perikarya of non-immunoreactive neurons of the infundibular nucleus. These results suggest that the central nervous system elaborates beta-endorphin or immunologically related peptides.

[Effect of various inhibitors on oxidative activity and ultrastructure of rat liver mitochondria]. / Influence de différents inhibiteurs sur l'activité d'oxydation et sur l'ultrastructure des mitochondries de foie de rat

Parallel studies concerning the influence of some inhibitors on electron transfer, and ultrastructural aspects of rat liver mitochondria led to the conclusion that metabolic activity and mitochondrial ultrastructure are closely linked ; when mitochondria oxidize succinate, their configuration change from the "condensed" to the "orthodox" state ; this change does not occur if the oxidation is inhibited.